PURIFICATION OF HUMAN COLONIC AND GASTRIC MAST-CELLS

We have developed a method to purify mast cells from enzymatic isolate s of human colonic mucosa (HCM) and submucosa/muscle (HCS), and gastri c mucosa (HGM) and submucosa/muscle (HGS). The purification of mast ce lls from these enzymatic isolates involves positive affinity-magnetic selection of mast cells using a monoclonal antibody specific for the c -kit receptor tyrosine kinase (CD117). The monoclonal antibody is coup led to Dynabeads for positive affinity selection of c-kit receptor pos itive cells which includes mast cells. This selection procedure genera tes preparations of mast cells from HCM, HCS, HGM and HGS that are 80% pure. The purified mast cells were microscopically normal and viable (> 85%). The functionality of purified mast cells was examined by stud ying the effect of anti-human IgE, Concanavalin A (Con A) and calcium ionophore A23187 on histamine release. These results show that this pu rification procedure generates microscopically normal, viable and func tional mast cells. This method of purifying human gastrointestinal tis sue mast cells may be a valuable tool for the further study of mast ce ll heterogeneity and the role of mast cells in the gastrointestinal tr act. (C) 1998 Published by Elsevier Science B.V. All rights reserved.